SUMMARY Cryo-electron microscopy (cryo-EM) has revolutionized imaging in biomedical science, from near atomic resolution visualization of individual biological molecules by single particle cryo-EM to the definition of macromolecular complexes, in situ, using cryo-electron tomography (cryo-ET). This revolution has been driven largely by advances in the detectors employed to collect the cryo-EM images, in the computer software programs utilized in image analysis and specimen reconstruction, and in the microscopes themselves. Because the technology underpinning cryo-EM is both expensive and has dramatically advanced over a short period of time, many research groups that could benefit from incorporating cryo-EM in their biomedical studies neither have access to cryo-EM technology nor have the training to employ the technology. To address access to cryo-EM technology and education, we will create the Pacific Northwest Center for Cryo-EM. The PNCC will be a collaborative effort between the Oregon Health and Science University (OHSU) and the Pacific Northwest National Laboratory (PNNL). By harnessing the strengths of OHSU in high resolution imaging, biological sample preparation and education in cryo-EM with the long-standing experience of PNNL in electron microscopy, high performance computing and robust user facility operation, we will endow the PNCC with outstanding capabilities in single particle cryo-EM and cryo-ET, as well as with a thorough and complete program in cryo-EM education or cross-training. Moreover, because the PNCC has established collaborations with leaders in cryo-EM technology and in computer hardware and software, the PNCC will be at the forefront of technological capability. Furthermore, the participation of OHSU in a cryo-EM school co-sponsored by the microscope manufacturer, FEI, will bolster the cross-training program of the PNCC. Both OHSU and PNNL will make major investments in the PNCC. OHSU will provide a low vibration space for the instrumentation, it will donate a new microscope to the center and OHSU will provide $19M in additional funding for use in space renovation, acquisition of an additional microscope and support of center staff. At the same time, PNNL has pledged access to their high performance computer centers, their user programs and their high resolution electron microscopes. By bringing together two major research institutions with complementary strengths, together with judiciously chosen collaborations and major additional sources of support, we will create a state- of-the-art, user-focused center for the high resolution imaging of biological molecules and macromolecular assemblies and for the education of the next generation of scientists trained in the field of cryo-EM.